Catgut Implantation At Acupoints For Allergic Rhinitis:a Double-Blind, Randomized, Sham-Controlled Trial And A Study On The Immune Mechanism In Rat Model

Object:To identify the clinical efficacy and safety and explore the mechanism of catgut implantation at acupoints for allergic rhinitis.Methods:1. This was a randomized, double-blind sham-controlled study, in which real versus sham catgut implantation at acupoints was compared in128patients with a history of PAR (persistent allergic rhinitis) or IAR (intermittent allergic rhinitis) and with a positive skin prick test (SPT). In the study, patients were randomly assigned by computer-generated randomization list into two groups and assessed prior to treatment. Then, they received two sessions of treatments (once per2weeks) and have a follow-up phase of8weeks. Each group was administred catgut implantation at the same acupoints:LI20, GB20, LI11, LI4and ST36. And the treatment was performed double-blindly by a well-trained physician in acupuncture. The main outcome measures included the primary and secondary indicators. Primary indicators were subjective symptoms scores evaluated by visual analogue scales (VAS) and Rhinoconjunctivitis Quality of Life Questionnaires (RQLQ). The secondary indicators were the results of endoscopic and laboratory examinations, such as the color of the nasal mucosa, the volume of the inferior turbinate, serum allergen-specific IgE, and some cytokines (IL-4, IL-10, IL-12, IFN-y and VCAM-1). Furthermore, the use of anti-allergic medication and adverse events were also recorded as one of the secondary indicators. The datum were recorded and analysed before the treatment, in the4th and8th week after the treatment to assess the efficacy and safety of catgut implantation at acupoints in treatment of AR.2. Thirty ovalbumin (OVA)-sensitized rats were randomly assigned into the study group or the control one. And another15rats were used as no-sensitized and no-treatment group. When rats in the study group had been treated with cautgut implantation at Yingxiang (LI20) for1sessions, the accumulative scores of allergic symptoms were assessed. The nasal mucosal pathological changes were studied by hematoxylin-eosinstaining (HE) staining. And the changes of neuropepides of the nasal mucosa, such as substance P(SP), calcitonin gene related peptide (CGRP) and neurokinin A (NKA), were studied by immunohistochemical and pathological methods. Then integrated optical densities of the neuropeptides were estimated.3. Nine rats were divdied into the study group (group A), the control (group B) or the blank control (group C) one. The rats in group A and B were sensitized in the same way as that of method2. After10days of the treatment of catgut implantation at Yingxiang (LI20), nasal mucosas of all the rats were collected. Then after the steps of RNA isolation, removal of contaminationg DNA from RNA preparations, RNA cleanup, and assessing RNA yield and quality, real-time PCR detections were performed by the Rat Inflammatory Response&Autoimmunity RT2ProfilerTM PCR Arrays. At last, the datum were analyzed by AACt methods.Results:1. The clinical efficacy and safety of catgut implantation at acupoints for allergic rhinitis1) The baselines of the two groups showed no statistical differences.2) Though the VAS sores for group A and group B were both significantly improved in the4th and8th week after the treatment (P<0.001) in self control study, only the improvements in the8th week after the treatment were clinical significant in inter group comparison (P<0.05). The changes in the VAS scores were of no significantly differences between the two groups in the4th week after treatment (P=0.753).3) The RQLQ socres for group A and group B were both significantly improved after the treatment (P<0.001), and the improvements in group A were significantly better than those in group B after treatment (P<0.05).4) For endoscopic examination, only in the8th week after treatment the changes of the color of nasal mocosas were significantly improved (P<0.05) in group A and group B, while the changes of the nasal secretion and the volume of the inferior turbinate were significantly improved in both of the4th and8th week after treatment in the two groups (P <0.01). And in inter group comparison, these two scoring items kept on significantly improving in group A (P<0.05), but there were no significant differences of the color of nasal mocusas between the groups in the whole course of assessment (P>0.200)5) The levels of serum specific IgE were significantly improved for both group A and group B after treatment (P<0.05), while there were no significant differences of other cytokines in neither of the groups (P>0.100). In inter group comparison, the differences of serum specific IgE were significant between the two groups either before or after the treatment. But the other cytokins were of no significant differences during the whole course (P>0.05).6) No adverse events occurred in group A or group B in the treatment.2. Regulatory Effect of Catgut Implantation at Acupoint on Neurogenic Inflammation in Rat Animal Model with Allergic Rhinitis1) Symptoms of allergic rhinitis were remarkably relieved after the treatment of catgut implantation at acupoints.2) Immunohistochemical study revealed a marked decrease of substance P (SP), neurokinin A (NKA) and calcitonin gene related peptide (CGRP) in the nasal mucosa of group A. Hematoxylin-eosinstaining (HE) staining demonstrated that edema of nasal mucosa was alleviated and the number of inflammatory cells decreased in group A. Vasodilation was significantly inhibited in nasal mucosa of group A.3. Studies on catgut implantation at acupoints for rat’s allergic rhinitis by RT2RNA QC PCR Arrays1) The quality of the RNA isolated from the nasal mucosal complied with the requirements.2) Most of the Ct data of the amplification curve were between20and35, and the curve inflection points were clear. Exponential phases were obvious especially in the samples of low concentration. None of the baselines moved ahead. And all of the curves kept parallel, which indicated the amplification efficiencies of the wells were similar. These results showed that the reaction system and PCR protocol were set reasonably.3) The melting curves of the PCR products were unimodal, which implied there was not any unspecific product appearing in the reaction.4) In the pair of group B and group A, there were2upregulated genes without statistical difference and17downregulated genes, in which10genes showed significant differences.5) When the data of group C were compared with that of group B,23genes were downregulated more than three times, but without statistical differences.6) There were44genes with a downregulated ratio of more than3times when the datum of group C were divided by those of group A. And9of the44genes showed statistical differences.Conclusion:1. Catgut implantation at acupoints is beneficial for alleviating the VAS and RQLQ scores. And because of the stimuli of the catgut imbedded in the acupoint, the improvement of the symptoms could last much longer when real versus sham catgut implantation at acupoints is compared. This treatment could also help to improve the color of mucosal, alleviate the nasal secretion and sewlling of inferior turbinate, and ameliorate the exprseeion of serum specific IgE. But it seems that catgut implantation is of no use for changing the cytokines involed in the immune reaction.2. Catgut implantation at acupoints could improve the symptoms of allergic rhinitis by regulating the neuropeptides of the nasal mucosa. But the neuropeptides keep on being statistical differential between the treatment group and the blank control one,while the rats treated by catugut implantation showed significant improvements of symptoms. This indicates inflammatory reaction of the nasal mocosa is still going on although the symptoms of allergic rhinitis are alleviated after this treatment.3. It is rational to use RT2ProfilerTM PCR Arrays to find changes of genes expression in the treatment of catgut implantation at acupoints for allergic rhinitis. Compared with the technicque of microarray gene expression, this method could screen the genes with differential expressions with a higher efficiency. And more proofs about the mechanism of catugut implantation at acupoints for allergic rhinitis are hoped to be provided with the development of technology.